146 results about "Liraglutide" patented technology

The invention discloses a method for preparing liraglutide by convergent synthesis. The method comprises the steps of performing solid phase synthesis to obtain four side chain protected peptide fragment sequences, gradually coupling the peptide fragments in a solution system to obtain an all-protected liraglutide straight chain polypeptide, removing the side chain protection of the 20th Lys, performing modification to form all-protected liraglutide, then cracking to remove protecting groups to obtain a crude liraglutide peptide, purifying and exchanging salt to obtain liraglutide; wherein in the four peptide fragment sequences, the first peptide fragment sequence is 1st to 8th amino acids in the liraglutide sequence, the second peptide fragment sequence is 9th to 16th amino acids in the liraglutide sequence, the third peptide fragment sequence is 17th to 26th amino acids in the liraglutide sequence, and the fourth peptide fragment sequence is 27th to 31st amino acids in the liraglutide sequence. By adopting the method, the yield is improved, and the synthesis cost is greatly reduced; and the method is favorable for large-scale and industrialized production.

The invention relates to a formula of a liraglutide sustained-release microsphere preparation. The preparation mainly comprises 5 to 60 parts of liraglutide and 50 to 200 parts of biodegradable substrate material, wherein the biodegradable substrate material is polylactic acid (PLA), polylactic acid-glycollic acid (PLGA) block copolymer, glycolide-lactide (PLCG) copolymer, polyglycolic acid (PGA), polycaprolactone (PCL), polylactic acid-glycollic acid-polycaprolactone (PLGA-PCL) copolymer or PLCG-polycaprolactone (PLCG-PCL) or a mixture of any two or more than two of the compounds, preferably the PLGA-PCL copolymer. The sustained-release microspheres have a high encapsulating rate and a good sustained-release effect.

The invention relates to the field of medicine synthesis, and discloses a method for synthesizing liraglutide. The method includes the steps that three polypeptide fragments of the first to the fourth amino acid, the fifteenth to the sixteenth amino acid and the seventeenth to thirty-first amino acid are synthesized at first according to the amino acid sequence from N end to C end of a main chain of liraglutide, and then the three polypeptide fragments and the other amino acid are connected according to the sequence from the C end to the N end in a coupling mode to synthesize the liraglutide. According to the method, the three fragments can be synthesized simultaneously, coupling and acidolysis with much carrier resin are avoided, the use quantity of resin carriers is greatly reduced, the synthesis cycle is shortened, and the complexity level of synthetic process is simplified on the premise that high total recovery and purity are guaranteed.

The invention belongs to the technical field of polypeptide preparation and relates to a preparation method of a liraglutide intermediate GLP-1(7-37). The preparation method utilizes a genetic recombination technology. Compared with chemical synthesis, the preparation method reduces impurities and improves purity and a yield. The preparation method comprises the following steps of introducing plasmids with hI-GLP-1(7-37) gene sequences into cells of escherichia coli by a gene engineering method to construct recombinant engineering bacteria, carrying out fermentation induction to obtain a hI-GLP-1(7-37) expression fusion protein, and carrying out renaturation, digestion conversion and separation purification to obtain GLP-1(7-37).

The invention relates to the biomedical field, and in particular relates to a method for efficiently expressing recombinant liraglutide. The method for efficiently expressing recombinant liraglutide is used for performing gene cloning and transformed expression. The recombinant liraglutide is constructed, and liraglutide protein is prepared through an escherichia coli expression technology. Compared with the two methods, the method disclosed by the invention is simple and convenient to operate, high in yield and low in cost.

A synthesis method for low-racemization impurity liraglutide comprises the following steps: performing synthesis to obtain a propeptide, coupling 2 to 5 peptides comprising Thr-Phe on the propeptide by using a solid-phase synthesis method; further, performing solid-phase synthesis to obtain a liraglutide resin; the liraglutide resin is cracked after modification, or the liraglutide resin is directly cracked, purified and frozen dry, so as to obtain the liraglutide. The provided liraglutide synthesis method effectively restrains or reduces the generation of racemization impurity D-Thr5 highly similar to a product property, which facilitates the purification of the coarse liraglutide, and the high yield of the liraglutide is ensured, thereby greatly reducing production costs; during the synthesis of the liraglutide, the syntheses between dipeptide fragments, tripeptide fragments, the tetrapeptide fragments and pentapeptide fragments and the Gly-resin or the syntheses between the combination of the dipeptide fragments, the tripeptide fragments, the tetrapeptide fragments and pentapeptide fragments and the Gly resin can be carried out at the same time, and accordingly the synthesis time is shortened to some extent.

The invention belongs to the technical field of preparation methods of polypeptides and particularly relates to a preparation method of liraglutide intermediate polypeptide GLP-1 (glucagon-like peptide-1) (7-37). The method comprises the following main steps: constructing recombinant liraglutide engineering bacteria, expressing liraglutide intermediate fusion protein in a form of an inclusion bodyunder Echerichia coli induction, and performing denaturation, renaturation, enzyme digestion, separation and purification to obtain the liraglutide intermediate polypeptide GLP-1 (7-37). Expression is changed into intracellular insoluble inclusion body expression by changing recombinant sequence signal peptides, and expression quantity is increased greatly; the washed inclusion body is subjectedto alkali dissolution, a large quantity of denaturant is not needed, the inclusion body with high concentration of protein concentration being 20-30 g / L is added to an inclusion body dissolution buffer, denaturation and renaturation time does not exceed 1 h, and enzyme digestion can be performed after dissolution; procedures are reduced, operation volume is reduced, reagent cost is reduced, and industrialized enlargement is facilitated; UniSP-50XS cation exchange is adopted for separation and purification, and separation degree is high. The purity of the liraglutide intermediate polypeptide prepared with the method reaches 87% or higher, and the yield is higher than 85%.

The invention belongs to the technical field of polypeptide preparation, and in particular, relates to a fusion protein and a method for preparing a liraglutide intermediate polypeptide GLP-1 (7-37).The method includes the main steps: constructing recombinant liraglutide intermediate engineering bacteria, expressing a GLP-1 (7-37) fusion protein by culture and induction of escherichia coli, and carrying out denaturation, renaturation, enzyme digestion and separated purification to obtain the intermediate polypeptide GLP-1 (7-37). Through changing a sequence of leading peptide, the expressionmode becomes an intracellular insoluble inclusion body expression, and the expression quantity is remarkably increased; inclusion bodies after washing are dissolved at high pH, and a large amount of denaturants are not needed to use, the inclusion bodies after washing are added to a inclusion body dissolved buffer solution at a high protein concentration of 5-40 g / L, the renaturation time does notexceed 1 h, and enterokinase digestion is carried out immediately after dissolution; the renaturation process can be reduced, the enzymatic digestion system is reduced, the cost of chemical reagentscan be reduced, and industrial amplification is facilitated; ion exchange separation and purification can be used, and the separation degree is high. The prepared liraglutide intermediate polypeptideGLP-1 (7-37) reaches 92% or more and the yield is more than 87%.

The invention relates to the field of polypeptide synthesis, and particularly relates to a preparation method of liraglutide. The preparation method comprises the following steps: coupling Gly and resin to obtain Gly-resin; carrying out primary successive coupling on the Gly-resin to obtain polypeptide segment-resin; and carrying out side chain modification, secondary successive coupling, cracking, purification and freeze-drying on the polypeptide segment-resin to obtain the liraglutide, wherein the sequence of the polypeptide segment in the polypeptide segment-resin is disclosed as SEQ ID NO:1, and the sequence of the liraglutide is disclosed as SEQ ID NO:2. The side chain modification process comprises the following steps: removing the protective group on the Lys side chain in the polypeptide segment-resin, and sequentially coupling Fmoc-Glu-OtBu and palmityl chloride. The liraglutide prepared by the preparation method has fewer impurities, is easy to purify and can enhance the yield of the final product.

The invention discloses a GLP-I analogue liraglutide sustained-release microspheres and a preparation method thereof. The GLP-I analogue liraglutide sustained-release microspheres comprise, by weight, 0.5 to 30% (w / w) of GLP-I analogue liraglutide, 70 to 99.5% of a polymer which has the molecular weight of 5000 to 300000 dalton, can be biodegraded and has biocompatibility, and 0 to 10% of pharmaceutically acceptable auxiliary materials. The GLP-I analogue liraglutide sustained-release microspheres have the average grain size of 5 to 40 micrometers and an encapsulation ratio more than 90%. The GLP-I analogue liraglutide sustained-release microspheres have a sustained-release period of several days or several months, obviously reduce use frequency, improve GLP-I analogue liraglutide bioavailability, reduce toxic and side effects and are conducive to clinical treatment.

The invention discloses a preparation method of liraglutide. The preparation method comprises the following steps: using a deprotection reagent Fmoc-Gly-wang resin for deprotection, and removing Fmoc protective groups; coupling three protective amino acids at the 12th-14th sites by protective peptide fragments X; coupling three protective amino acids at 20-22 sites by protective peptide fragments Y; coupling protective amino acid at the 31th site by a Boc-His(trt)-OH protective form; removing alloc protection of lysine side chains for main-chain peptide resin of the liraglutide, coupling the side chains one by one and thus obtaining peptide resin of the liraglutide; cracking the peptide resin of the liraglutide in cracking liquid and thus obtaining a liraglutide crude product; purifying the liraglutide crude product and thus obtaining a liraglutide fine product. The preparation method disclosed by the invention has the advantages that segment coupling is carried out on amino sites which are easiest for folding in the process of synthesizing the main chain of the liraglutide, so that the problems of multiple times of coupling and low access rate for the amino acids due to folding in the synthesis process are avoided, the production cycle of the liraglutide is shortened, and the synthetic yield of the liraglutide is increased.